Anticipating and strategizing to address potential bottlenecks during clinical research projects in sub-Saharan Africa: a case for adapting approaches and tools used in the entrepreneurial and development sectors.

Many research funders have invested billions of US dollars in building research capacity in sub-Saharan Africa (SSA). Despite these colossal investments, many well-intentioned and designed clinical research projects have either failed to kick off or ended abruptly. Although obstacles to clinical research in SSA are well known, there is limited information on frameworks and tools that can be used to anticipate and avert these systemic bottlenecks, particularly those related to socio-politics. In this paper, we leveraged lessons from entrepreneurs and development experts in harsh and uncertain business environments to develop a framework for anticipating and addressing potential bottlenecks to clinical research in SSA. More so, to illustrate and build a case for this framework, we shared our experience in supporting clinicians and regulators to adopt a point-of-use care tool, the "chemoPAD," to screen for the quality of anticancer medications rapidly and systematically in Cameroon despite resistance from some stakeholders. The critical steps in this framework involve identifying stakeholders, categorizing them based on their potential reactions to the study (adversary, supporters, and indifferents), and developing critical strategies to engage or deal with each stakeholder's reactions, starting with adversaries. This approach may be useful in complex research projects, especially clinical trials, which often involve many stakeholders with different interests and perceptions.

Noninvasive Approach of Plasmodium falciparum Molecular Detection for Malaria Surveillance in Malaria Endemic Areas in Cameroon.

Background: Accurate, cost-effective, and noninvasive alternative molecular methods are needed for detecting low malaria parasitemia. The currently-used nested polymerase chain reaction (nPCR) requires blood as well as skilled personnel in order to minimise the risk of bloodborne disease transmission. Therefore, this study is aimed at assessing the accuracy of a noninvasive and more affordable malaria diagnosis with saliva using the loop-mediated isothermal amplification (LAMP) technique. Methods: A cross-sectional study was conducted in the Centre and Southwest regions of Cameroon. Matched blood and saliva samples collected from symptomatic and asymptomatic participants were tested for malaria using rapid diagnostic tests, microscopy, PCR, and LAMP. Statistics were performed using R studio software at 95% confidence interval. Results: A total of 100 participants (65% symptomatic and 35% asymptomatic) aged between 1 and 74 years with a balanced gender distribution ratio of 1.08 were included in our study. The prevalence of malaria was 61%, 57%, 59%, 42%, 35%, 17%, and 16% for blood-RDT, blood-PCR, blood-LAMP, blood-RT-LAMP, saliva-PCR, saliva-RT-LAMP, and saliva-LAMP, respectively. Both saliva and blood showed a sensitivity of 43.90% and respective specificities of 68.75% and 57.62%. When using RT-LAMP, sensitivities of 49.38% and 48.21% and specificities of 94.11% and 66.67% were recorded for saliva and blood, respectively. Sensitivities of 70.23% and 73.49% and specificities of 62.5% and 76.47% were recorded, respectively, for saliva-LAMP and saliva-RT-LAMP when compared to saliva-PCR as the gold standard. Saliva-LAMP and saliva-RT-LAMP had a fair agreement (к = 0.221 and 0.352, respectively) with saliva-PCR. Homemade LAMP and RT-LAMP technologies match the WHO recommendations and after proper validation in a larger sample size, could serve for malaria diagnosis in developing countries.

The impact of COVID-19 on implementation of mass testing, treatment and tracking of malaria in rural communities in Ghana: A qualitative study.

BACKGROUND: Mass test, treat and track (MTTT) of malaria is ongoing in the Pakro sub district of Ghana. In the delivery of MTTT of malaria, community health volunteers are trained to routinely provide this service through a door-to-door strategy. Following the report of the first cases of COVID-19 in Ghana, we conducted this study to explore the effects of the pandemic on the implementation of the MTTT of malaria intervention. METHODS: Using qualitative methodology, we conducted ten focus groups discussions (FGDs) in eight communities: eight with community members (N = 49); one with health workers (N = 6), and one with MTTT of malaria volunteers. In addition, two in-depth interviews (IDI) were conducted, one with health worker and another with a health manager. All interviews were recorded, translated into English during transcription and analysed using QSR NVivo 12. Thematic content analysis was used in this study. RESULTS: The findings of the study showed an increase in the number of people reporting with complications of malaria in health facilities in the study communities during the COVID-19 period. Some participants were of the view that COVID-19 rumours and misinformation could largely be responsible for the low coverage and uptake of the MTTT of malaria intervention. To sustain the uptake of the MTTT intervention, community engagement strategies were employed to identify and respond to these rumours. Also, incentive schemes were introduced to encourage parents and children to participate in the MTTT intervention during this period of COVID-19. CONCLUSION: Findings suggest that the COVID-19 pandemic has adversely affected the provision and uptake of malaria prevention and treatment services, especially the MTTT of malaria being implemented at the community level. These observations underscore the need to find innovative ways to address the challenges encountered in providing essential services during public health emergencies.

Drug resistance markers within an evolving efficacy of anti-malarial drugs in Cameroon: a systematic review and meta-analysis (1998-2020).

BACKGROUND: Malaria remains highly endemic in Cameroon. The rapid emergence and spread of drug resistance was responsible for the change from monotherapies to artemisinin-based combinations. This systematic review and meta-analysis aimed to determine the prevalence and distribution of Plasmodium falciparum drug resistance markers within an evolving efficacy of anti-malarial drugs in Cameroon from January 1998 to August 2020. METHODS: The PRISMA-P and PRISMA statements were adopted in the inclusion of studies on single nucleotide polymorphisms (SNPs) of P. falciparum anti-malarial drug resistance genes (Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, Pfatp6, Pfcytb and Pfk13). The heterogeneity of the included studies was evaluated using the Cochran's Q and I2 statistics. The random effects model was used as standard in the determination of heterogeneity between studies. RESULTS: Out of the 902 records screened, 48 studies were included in this aggregated meta-analysis of molecular data. A total of 18,706 SNPs of the anti-malarial drug resistance genes were genotyped from 47,382 samples which yielded a pooled prevalence of 35.4% (95% CI 29.1-42.3%). Between 1998 and 2020, there was significant decline (P < 0.0001 for all) in key mutants including Pfcrt 76 T (79.9%-43.0%), Pfmdr1 86Y (82.7%-30.5%), Pfdhfr 51I (72.2%-66.9%), Pfdhfr 59R (76.5%-67.8%), Pfdhfr 108 N (80.8%-67.6%). The only exception was Pfdhps 437G which increased over time (30.4%-46.9%, P < 0.0001) and Pfdhps 540E that remained largely unchanged (0.0%-0.4%, P = 0.201). Exploring mutant haplotypes, the study observed a significant increase in the prevalence of Pfcrt CVIET mixed quintuple haplotype from 57.1% in 1998 to 57.9% in 2020 (P < 0.0001). In addition, within the same study period, there was no significant change in the triple Pfdhfr IRN mutant haplotype (66.2% to 67.3%, P = 0.427). The Pfk13 amino acid polymorphisms associated with artemisinin resistance were not detected. CONCLUSIONS: This review reported an overall decline in the prevalence of P. falciparum gene mutations conferring resistance to 4-aminoquinolines and amino alcohols for a period over two decades. Resistance to artemisinins measured by the presence of SNPs in the Pfk13 gene does not seem to be a problem in Cameroon. Systematic review registration PROSPERO CRD42020162620.

[Frequency and titration of hemolytic activity of anti-A and anti-B antibodies in mothers of children with jaundice in Yaoundé, Cameroon]. / Fréquence et titrage des hémolysines anti-A et anti-B chez les mères d'enfants ictériques à Yaoundé, Cameroun.

INTRODUCTION: The alloimmunization of the ABO blood group system is involved in neonatal jaundice with a considerable overall prevalence. The role of ABO incompatibility is relatively little known. The purpose of this study was to investigate neonatal jaundice due to feto-maternal ABO incompatibilities and to determine the link between the hemolysins value in the mother and the degree of jaundice observed in the infant. METHODS: We conducted a cross-sectional study from June to November 2015. The study population was exclusively composed of moms who were blood type O with children who were a different blood type hospitalized in the Department of Neonatology at the Reference Hospital in the city of Yaoundé. Statistical analyses were performed using the GraphPadPrism 6 software with a confidence interval of 95%. RESULTS: Hemolysins frequency was of 20.58% (7/34) and anti-A hemolysin was the most common type (85.7%; 6/7). The new-born who had blood type B had a greater concentration of bilirubin levels compared to those of the AB group (p = 0.01). Multiparity was not associated with the presence of hemolysin (p = 0.8) as well as blood type of the infant was not associated with the occurrence of the hemolysins in the mother (p = 0.5). CONCLUSION: Early neonatal jaundice or protracted neonatal jaundice are also caused by hemolysins anti-A and anti-B derived from the allo-ABO immunization. A study on a larger sample is recommended for better assessment.

Negative Association of Interleukin-33 Plasma Levels and Schistosomiasis Infection in a Site of Polyparasitism in Rural Cameroon.

Background: This study aimed to investigate the association of plasma levels of IL-33, a mucosal alarmin known to elicit type-2 immunity, with infection and liver fibrosis profiles of school children from an endemic area for Schistosoma mansoni, malaria and hepatitis (B & C) in rural Cameroon. Methods: A cross-sectional study enrolling schoolchildren from 5 public schools was conducted. Single schistosomiasis, malaria and hepatitis infections or co-infections were assessed by kato katz, microscopy, and rapid diagnostic tests, respectively. Hepatic fibrosis was assessed by ultrasound according to WHO Niamey guidelines and plasma levels of Interleukin 33 were determined by ELISA. All statistics were performed using R studio software. Principal findings: We found a prevalence of 13.5% (37/275), 18.2% (50/275), and 8% (22/275), respectively for schistosomiasis, malaria and hepatitis (B or C) single infections. Only 7.6% (21/275) of co-infections were reported. Although Plasma IL-33 showed a minimal negative risk for schistosomiasis infection (AOR 0.99; 95% CI 0.97-1.01), S. mansoni infected participants had lower levels of plasma IL-33 (p = 0.003) which decreased significantly as eggs burdens increased (p = 0.01) with a negative Pearson coefficient of r = -0.22. Hepatic fibrosis occurred in 47.3% (130/275) of our study population independently from plasma levels of IL-33 (AOR 1.00; 95% CI 0.99-1.01). Conclusion/Significance: Our data failed to show an association between plasma IL-33 levels and liver disease but convincingly report on a negative association between plasma IL-33 levels and schistosomiasis infection and egg burden in school children from a polyparasitic schistosomiasis endemic area.

Schistosomiasis Burden and Its Association With Lower Measles Vaccine Responses in School Children From Rural Cameroon.

Background and Methods: Schistosomiasis is debilitating and reported to impair immune responsiveness of infected hosts. In Cameroon, mass drug administration (MDA) is used in schoolchildren to reduce transmission of S. haematobium and S. mansoni. The effects of MDA and the impact of schistosomiasis on the titers of antibodies in vaccinated children have been poorly studied. We therefore assessed the prevalence of schistosomiasis in schoolchildren, eight months after MDA, in two locations: Barombi Koto (BK), endemic for S. haematobium (N = 169) and Yoro (Y), endemic for S. mansoni (N = 356). Age, gender, residence time and frequency of contact with river water were assessed as risk factors for infection and morbidity in both localities. In 70 schoolchildren from BK and 83 from Y, ultrasound was used to assess morbidity according to the WHO guidelines. Evaluation of measles antibodies was performed in previously vaccinated schoolchildren (14 with S. haematobium and 12 egg-negative controls from BK and 47 with S. mansoni and12 egg-negative controls from Y). Principal Findings and conclusions: The prevalence of S. haematobium was 25. 4% in BK (43/169) and 34.8% for S. mansoni in Y (124/356), indicating the persistent transmission of schistosomiasis despite MDA. Older age (AOR 1.31; 95%CI 1.12-1.54) and higher frequencies of exposure to river water (AOR 1.99; 95%CI 1.03-3.86) were identified as risks for infection in BK whereas only older age (OR 1.15; 95%CI 1.04-1.27) was a risk for infection in Y. Bladder pathology (score 2 to 5) was observed in 29.2% (7/24) of egg-positive children in BK and liver pathology (pattern C) in 31.1% (19/61) of egg-positive children in Y. There was a positive correlation between S. haematobium egg burden and bladder pathology (AOR 1.01; 95% CI 0.99-1.02) and positive correlation between S. mansoni-driven liver pathology and female gender (AOR 3.01; 95% CI 0.88-10.26). Anti-measles antibodies in vaccinated children were significantly lower in S. mansoni-infected when compared to egg-negative controls (p = 0.001), which was not observed in the S. haematobium-infected group from BK. Our results demonstrate a questionable efficacy of MDA alone in halting schistosomiasis transmission and confirm a possible immunomodulatory effect of S. mansoni on response to vaccines.

Host candidate gene polymorphisms and associated clearance of P. falciparum amodiaquine and fansidar resistance mutants in children less than 5 years in Cameroon.

BACKGROUND: In this post-hoc analysis, we determined the influence of single nucleotide polymorphisms in host candidate immune genes on the outcome of drug resistant malaria in Cameroon. METHODS: Human DNA from 760 patients from a previous clinical trial was subjected to mass spectrometry-based single nucleotide polymorphism (SNP) genotyping. Allele frequencies of candidate immune genes were calculated for 62 SNPs on 17 human chromosomes for their possible involvement in clearance of drug-resistant parasites with the triple mutations of pfcrt76T, pfmdr86Y, and pfmdr1246Y (TY) and pfdhfr51I, pfdhfr59R, pfdhfr108N, and pfdhps437G (IRNG) which were determined by dotblot or PCR-restriction analysis. Differences in SNP frequencies and association analysis were carried out by comparing Chi-square odds ratios (ORs) and stratified by Mantel-Haenzel statistics. An adjusted P value (OR) <0·0008 was considered significant. RESULTS: Post-treatment drug failure rates were amodiaquine (36·4%); sulpadoxine/pyrimethamine-amodiaquine combination (15·4%); and sulphadoxine/pyrimethamine (18·1%). SNPs in IL22, IL-4R1, and CD36 appeared to have been associated with clearance of resistant parasites [p  =  0·017, OR (C allele):1·44, 95% CI (OR): 1·06-1·95]; [P  =  0·014, OR  =  1·31, 95% CI (OR): 1·07-1·83]; [P  =  5·78×10(-5), OR  =  0·27, 95%CI (OR): 0·13-0·54], respectively, with high fever (>39°C for 48 hours) [IL-22, P  =  0·01, OR  =  1·5, 95% CI (OR): 1·8-2·1] and also in high frequency among the Fulani participants [P  =  0·006, OR  =  1·83, 95% CI (OR): 1·11-3·08)]. The CD36-1264 null allele was completely absent in the northern population. CONCLUSION: Independent association of SNPs in IL22 and IL-4 with clearance of amodiaquine- and sulphadoxine/pyrimethamine-resistant parasites did not reach statistical significance, but may suggest that not all drug-resistant mutants are adversely affected by the same immune-mediated mechanisms of clearance.